LOCUS       sequence-449187-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 IPTG-driven Cas13a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  AUTHORS   Fishman CB, Crawford KD, Bhattarai-Kline S, Poola D, Zhang K, 
            Gonzalez-Delgado A, Rojas-Montero M, Shipman SL
  TITLE     Continuous multiplexed phage genome editing using recombitrons.
  JOURNAL   Nat Biotechnol. 2024 Sep 5. doi: 10.1038/s41587-024-02370-5.
  PUBMED    39237706
REFERENCE   2  (bases 1 to 6890)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2024 Sep 5. doi: 10.1038/s41587-024-02370-5."
COMMENT     Sequence Label: pKDC.118
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(3557..3575)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3571..3618
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3789..4580)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4581..4672)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4720..5458)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5668..6750)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6712..6731
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6751..6828)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6874..6890,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6874..6890,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgaaagtga cgaaggtagg aggcatttcg cataagaagt acacgtccga
      121 aggccgctta gtgaagtcag aatcggaaga aaatcgcaca gacgaacgtc tgtcggcgtt
      181 gcttaatatg cgccttgaca tgtatatcaa gaatcccagc agcacggaaa ccaaggaaaa
      241 tcaaaaacgc attgggaaat taaagaaatt cttctcaaac aaaatggtct atcttaaaga
      301 caataccttg agtttgaaga atgggaaaaa ggagaacatt gatcgtgagt attctgagac
      361 tgacatcctt gagagcgatg tccgtgacaa gaaaaacttc gccgtgttga aaaagatcta
      421 tctgaatgaa aacgtgaact cggaggaatt ggaagttttt cgtaacgaca ttaagaagaa
      481 actgaacaaa atcaacagcc tgaagtactc atttgaaaag aataaggcga attatcaaaa
      541 gattaatgag aataacatcg agaaggttga aggtaagtca aagcgtaaca ttatttacga
      601 ttattatcgt gagtcagcga aacgtgacgc ttatgtaagc aatgtgaaag aagcctttga
      661 taagctttac aaggaagagg acattgcaaa acttgttctt gaaattgaga accttacgaa
      721 gttagagaaa tacaagattc gcgagttcta ccacgaaatt attggacgta agaatgacaa
      781 ggaaaacttt gcaaaaatca tctacgaaga aatccagaat gttaataaca tgaaagagtt
      841 gatcgagaag gtaccggaca tgagtgagtt gaaaaagagc caagtatttt acaagtatta
      901 cttagacaaa gaagagttga acgacaagaa catcaaatac gcgttttgtc atttcgtgga
      961 aatcgaaatg agtcagttgc tgaagaacta cgtatataag cgcttaagta atatctcgaa
     1021 tgacaaaatt aagcgtatct ttgaatacca gaacttgaaa aaattgatcg aaaataagct
     1081 gttaaacaaa cttgacacgt acgtccgtaa ttgtggaaag tataattatt atttgcaaga
     1141 cggcgaaatt gccacttcag atttcatcgc ccgcaaccgt cagaatgaag cgtttcttcg
     1201 caacatcatt ggggtgtcat ctgtggccta cttttctctt cgcaacattc ttgaaacgga
     1261 gaacgagaat gatattactg ggcgtatgcg cggcaaaaca gttaagaaca ataaaggtga
     1321 agagaagtac gtgtccggag aagttgataa gatctataat gaaaataaga agaacgaggt
     1381 taaggagaac ttaaaaatgt tctattcgta cgatttcaat atggacaaca agaatgaaat
     1441 cgaagatttc ttcgccaaca tcgacgaggc gatttcttcc atccgtcacg gtattgtcca
     1501 cttcaacttg gaattagaag gtaaggatat ctttgcgttc aagaacattg cgccatccga
     1561 aatctcaaag aagatgtttc agaatgagat taacgagaaa aaactgaaat tgaagatctt
     1621 tcgtcaactg aactctgcca acgtgttccg ctatctcgaa aagtataaaa ttctgaatta
     1681 ccttaaacgt acacgcttcg agtttgtcaa taaaaatatc ccattcgtcc cgtctttcac
     1741 caaattatat tcgcgcattg atgacctgaa gaatagtctt gggatttact ggaaaactcc
     1801 gaaaacaaac gacgacaata agactaagga gattattgat gcccaaatct atttgcttaa
     1861 aaacatctat tacggggagt tcctgaatta tttcatgtcg aacaatggta atttctttga
     1921 gatttctaaa gaaatcatcg aattgaacaa gaacgataaa cgcaacttaa agactgggtt
     1981 ttacaagctg caaaagtttg aagacatcca ggagaagatt ccaaaggaat acttggcgaa
     2041 tatccagtcc ctgtacatga ttaatgccgg taatcaggac gaagaagaaa aggacactta
     2101 tattgatttc attcaaaaga tcttcttaaa gggatttatg acgtatcttg ctaataacgg
     2161 tcgtttaagt ctgatttaca tcggctcgga tgaagaaaca aatacgtcat tagcagaaaa
     2221 gaagcaagag tttgacaagt tcttgaagaa gtacgagcag aacaataata tcaagatccc
     2281 ctatgagatc aatgaattcc tgcgtgagat caaactggga aacatcctga agtatactga
     2341 gcgtttaaac atgttctacc ttatcttaaa gcttttgaat cacaaggagc tgacaaatct
     2401 gaagggtagt cttgaaaaat atcagtctgc caataaggaa gaagcgttct ctgaccaatt
     2461 ggagttaatt aacctgctta accttgacaa caaccgcgtg acggaagact tcgaattaga
     2521 ggccgacgag attggaaaat ttcttgattt caatggcaac aaagttaagg ataacaagga
     2581 actgaaaaag ttcgatacaa acaagatcta ctttgacggc gagaacatta tcaaacaccg
     2641 tgccttctac aatattaaga aatatggcat gttaaactta ctggagaaaa ttgccgacaa
     2701 ggctggatac aagatctcga tcgaagagct gaagaaatac tccaataaaa agaatgagat
     2761 cgagaagaac cataagatgc aggaaaatct gcaccgcaaa tacgctcgtc cccgtaaaga
     2821 cgagaagttt acagatgagg actatgaaag ttacaagcaa gctattgaga atattgagga
     2881 gtacacccac cttaagaaca aggtagaatt caatgagctg aatttactgc agggcctgtt
     2941 gctgcgcatt ttacatcgtt tagtcggata tacctcaatt tgggaacgcg atctgcgctt
     3001 ccgccttaaa ggtgagttcc cagaaaacca atacatcgaa gagatcttca actttgaaaa
     3061 taagaagaac gtgaagtaca aagggggtca gattgtagag aaatacatta aattctacaa
     3121 ggaattacat caaaatgatg aagttaagat caacaagtac agttccgcga atatcaaggt
     3181 gttgaagcaa gaaaagaagg acctttatat tcgcaattac atcgcccact tcaattatat
     3241 tcctcacgcc gagatctcac tgctggaagt ccttgaaaat ttgcgtaaat tgctgtccta
     3301 cgatcgcaaa ctgaaaaatg ccgtaatgaa atcagtagtt gatatcctta aggagtatgg
     3361 ttttgtagcc acattcaaaa tcggggcgga caagaagatc ggtattcaga cactggagag
     3421 cgaaaaaatc gtgcatctta agaatcttaa gaagaagaag ttaatgactg accgcaattc
     3481 cgaggaactt tgcaaattgg tgaagattat gtttgaatac aaaatggaag agaaaaagtc
     3541 tgaaaacctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     3601 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt
     3661 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     3721 cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt
     3781 gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
     3841 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
     3901 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
     3961 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
     4021 actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
     4081 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
     4141 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
     4201 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
     4261 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
     4321 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
     4381 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     4441 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
     4501 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     4561 gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc
     4681 tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag
     4741 ttacccacag attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg
     4801 cgccggtggc gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct
     4861 tttccggtgc atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa
     4921 cccgacagga cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct
     4981 gttccgaccc tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg
     5041 cgctttctca tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct
     5101 gggctgtaag caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc
     5161 acttgagtcc aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg
     5221 ggagttcgca gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag
     5281 tccggctaca ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg
     5341 ttaagcagtt ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc
     5401 gtttacaggg caaaagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5461 ttctactgaa ccgctctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     5521 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     5581 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     5641 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5701 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5761 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5821 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     5881 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     5941 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     6001 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     6061 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     6121 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     6181 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     6241 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     6301 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     6361 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     6421 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     6481 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     6541 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     6601 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6661 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6721 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6781 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6841 gctctccctt atgcgactcc tgcattagga aattaatacg actcactata
//
